Spectrophotometric Method for Quantitative Determination of Total Anthocyanins and Quality Characteristics of Roselle (Hibiscus sabdariffa)

Abstract

A simple, rapid and cost-saving method for the determination of total anthocyanins in roselle has been developed. The method was based on pH-differential spectrophotometry. The calibration curve of the major anthocyanin in roselle, delphinidin 3-sambubioside (Dp-3-sam), was constructed by using methyl orange and their correlation factor. The reliability of this developed method was comparable to the direct method using standard Dp-3-sam and the HPLC method. Quality characteristics of roselle produced in Thailand were also reported. Its physical quality met the required specifications. The overall chemical quality was herein surveyed for the first time and it was found to be the important parameter corresponded to the commercial grading of roselle. Total contents of anthocyanins and phenolics were proportional to the antiradical capacity.

Key words

Hibiscus sabdariffa - roselle - Malvaceae - anthocyanins - pH-differential spectrophotometry - method development - validation - quality

References

• 1 Bunyapraphatsara N, Chansrakaew W, Chayamarit K, Chokechaijaroenporn O, Chuokul W. Hibiscus sabdariffa Linn. In: Farnsworth NR, Bunyapraphatsara N, editors Thai medicinal plants recommended for primary health care system. Bangkok; Prachachon 1992: 163-6.
• 2 Delgado-Vargas F, Paredes-Lopez O. Natural colorants for food and nutraceutical uses. Boca Raton; CRC Press LLC 2002: 257-98.
• 3 Ali B H, Wabel N A, Blunden G. Phytochemical, pharmacological and toxicological aspects of Hibiscus sabdariffa L.: A review.  Phytother Res. 2005;  19 369-75.
• 4 Tseng T H, Wang C J, Kao E S, Chu H Y. Hibiscus protocatechuic acid protects against oxidative damage induced by tert-butyl hydroperoxide in rat primary hepatocytes.  Chem Biol Interact. 1996;  101 137-48.
• 5 Wong P -K, Yusof S, Ghazali H M, Man Y BC. Physico-chemical characteristics of roselle (Hibiscus sabdariffa L.)  Nutr Food Sci. 2002;  32 68-73.
• 6 Du C T, Francis F J. Anthocyanins of roselle (Hibiscus sabdariffa L.)  J Food Sci. 1973;  38 810-2.
• 7 Wang C J, Wang J M, Lin W L, Chu C Y, Chou F P, Tseng T H. Protective effect of hibiscus anthocyanins against tert-butyl hydroperoxide-induced hepatic toxicity in rats.  Food Chem Toxicol. 2000;  38 411-6.
• 8 Delgado-Vargas F, Paredes-Lopez O. Natural colorants for food and nutraceutical uses. Boca Raton; CRC Press 2002: 167-211.
• 9 Petri G, Krawczyk U, Kéry Á. Spectrophotometric and chromatographic investigation of bilberry anthocyanins for quantification purposes.  Microchem J. 1997;  55 12-23.
• 10 Soriano A, Pérez-Juan P M, Vicario A, González J M, Pérez-Coello M S. Determination of anthocyanins in red wine using a newly developed method based on fourier transform infrared spectroscopy.  Food Chem. 2007;  104 1295-303.
• 11 Sáenz-López R, Fernández-Zurbano P, Tena M T. Development and validation of a capillary zone electrophoresis method for the quantitative determination of anthocyanins in wine.  J Chromatogr A. 2003;  990 247-58.
• 12 Mondello L, Cotroneo A, Errante G, Dugo G, Dugo P. Determination of anthocyanins in blood orange juices by HPLC analysis.  J Pharm Biomed Anal. 2000;  23 191-5.
• 13 Ancos B D, Ibanez E, Reglero G, Cano M P. Frozen storage effects on anthocyanins and volatile compounds of raspberry fruit.  J Agric Food Chem. 2000;  48 873-9.
• 14 Lee J, Durst R W, Wrolstad R E. Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: Collaborative study.  J AOAC Int. 2005;  88 1269-78.
• 15 Mazza G, Cacace J E, Kay C D. Methods of analysis for anthocyanins in plants and biological fluids.  J AOAC Int. 2004;  87 129-45.
• 16 Giusti M M, Wrolstad R E. Characterization and measurement of anthocyanins by uv-visible spectroscopy. Available at http://www.does.org/masterli/facsample.htm. Accessed July 7, 2003. 
• 17 Wrolstad R E, Durst R W, Giusti M M, Rodriguez-Saona L E. Analysis of anthocyanins in nutraceuticals. In: Ho C-T, Zheng QY, editors Quality management of nutraceuticals. Washington, DC; American Chemical Society 2001: 42-62.
• 18 European Pharmacopoeia 4.4 Nordlingen; Druckerei C. H. Beck 2001.
• 19 Validation of analytical procedures. Available at http://www.nihs. go.jp/drug/validation/q2bwww.html. Accessed August 23, 2005. 
• 20 Market survey: Hibiscus sabdariffa. Available at http://www.herbs. org/africa/hibiscus.html. Accessed December 12, 2006. 
• 21 Pale E, Koiuda-Bonafos M, Nacro M. Characterisation and antioxidative scavenging activities of anthocyanes plants of Burkina Faso.  Cr Chim. 2004;  7 973-80.
• 22 Plant dry ash/acid extraction for Ca, K, and Mg. Available at http://www.uga.edu/∼sisbl/plantash.html. Accessed January 3, 2007. 
• 23 Waterman P G, Mole S. Analysis of phenolic plant metabolites. Oxford and Northampton; Alden Press Limited 1994: 66-103.
• 24 Thai Herbal Pharmacopoeia. Bangkok; Prachachon 1995.
• 25 Leong L P, Shui G. An investigation of antioxidant capacity of fruits in Singapore markets.  Food Chem. 2002;  76 69-75.
• 26 Senese F. What is methyl orange? How is it made? Available at http://antoine.frostburg.edu/chem/senese/101/acidbase/faq/methyl-orange. shtml. Accessed January 3, 2007.